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首页> 外文期刊>Vaccine >Protection against dengue type 2 virus induced in mice immunized with a DNA plasmid encoding the non-structural 1 (NS1) gene fused to the tissue plasminogen activator signal sequence.
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Protection against dengue type 2 virus induced in mice immunized with a DNA plasmid encoding the non-structural 1 (NS1) gene fused to the tissue plasminogen activator signal sequence.

机译:在用编码与组织纤溶酶原激活物信号序列融合的非结构性1(NS1)基因的DNA质粒免疫的小鼠中诱导的登革热2型病毒的防护。

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摘要

Dengue is one of the most important arboviral diseases in humans, and although efforts over the last decades have dealt with the development of a vaccine, this vaccine is not available yet. In order to evaluate the potential of a DNA vaccine based on the non-structural 1 (NS1) protein against dengue virus (DENV), we constructed the pcTPANS1 plasmid which contains the secretory signal sequence derived from human tissue plasminogen activator (t-PA) fused to the full length of the DENV-2 NS1 gene. Results indicate that pcTPANS1 promotes correct expression of NS1 in eukaryotic cells and drives secretion of the recombinant protein to the surrounding medium in a dimeric form. Balb/c mice, intramuscularly inoculated with this plasmid, presented high levels of antibodies, recognizing mainly surface-exposed conformational epitopes present in the NS1 protein expressed by insect cells. Long-term antibody response was observed in animals 56 weeks after the first plasmid inoculation, and a rapid, efficient secondary response was observed after a DNA boost. Vaccinated animals were challenged against DENV-2 in two murine models, based on intracerebral (i.c.) and intraperitoneal (i.p.) virus inoculations, and in both cases, pcTPANS1-immunized mice were protected. Overall, these results provide further support for the use of such a plasmid in a possible approach for the development of a vaccine against DENV..
机译:登革热是人类最重要的虫媒病毒疾病之一,尽管过去几十年来人们一直在努力开发疫苗,但目前尚无这种疫苗。为了评估基于非结构性1(NS1)蛋白的DNA疫苗针对登革热病毒(DENV)的潜力,我们构建了pcTPANS1质粒,该质粒包含源自人组织纤溶酶原激活物(t-PA)的分泌信号序列与DENV-2 NS1基因的全长融合。结果表明pcTPANS1促进真核细胞中NS1的正确表达,并以二聚体形式驱动重组蛋白向周围培养基的分泌。用该质粒肌肉内接种的Balb / c小鼠表现出高水平的抗体,主要识别昆虫细胞表达的NS1蛋白中存在的表面暴露构象表位。第一次质粒接种后56周,在动物中观察到长期抗体反应,而在DNA加强免疫后观察到快速,有效的二次反应。基于脑内(i.c.)和腹膜内(i.p.)病毒接种,在两种鼠模型中对疫苗接种的动物进行了DENV-2攻击,在这两种情况下,都对pcTPANS1免疫的小鼠进行了保护。总体而言,这些结果为这种质粒在开发抗DENV疫苗的可能方法中的使用提供了进一步的支持。

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